Discovery and Development of New Enzymes for Biocatalysis in Organic Synthesis

The creation and evolution of new enzymes for synthesis is an exciting new area of research. A very low initial activity is all that is needed as a starting point to evolve new non-natural biocatalysts. The availability of a suitable high throughput screening method is essential for directed evolution of enzymes. The Carnell group has recently developed a colorimetric solid phase screen for the enzyme aldehyde oxidase (PaoABC) enabling us to screen several thousand mutants at a time. In this project we plan to exploit our novel plate screen to evolve new enzyme reactions by mutation of PaoABC. This enzyme is part of a family called xanthine oxidoreductases (XOR) that use the same catalytic chemistry for a spectrum of substrates including aldehydes, purines and imines. Our intention is to develop these enzymes and integrate them into synthetic enzyme cascades for the synthesis of bioactive molecules, for biomass conversion and for enantioselective oxidation of aldehydes.